Identification of DNA-Damage DNA-Binding Protein 1 as a Conditional Essential Factor for Cytomegalovirus Replication in Interferon-γ-Stimulated Cells
Figure 2
pM27 affects STAT2 protein levels.
(A) STAT2-HA cells were infected with indicated MCMV mutants (30 PFU/cell). Cells were incubated with or without cycloheximide (100 µg/ml) and actinomycin D (5 µg/ml). After 1, 2 or 5 h cells were analyzed by western blotting. The inhibitors precluded expression of pp89-IE1 and pM27-HA indicating tight inhibition. (B) 35S-L-Cys/L-Met labelled STAT2-HA cells were infected (5 PFU/cell, 20 h). Cells were metabolically labelled (30 min) and subsequently chased for the indicated time. STAT2-HA was precipitated using an HA-specific antibody. Proteins were separated by 8% SDS-PAGE and visualized by autoradiography. (C) As in (B), but cells were infected (20 PFU/cell) for 90 min prior to the indicated pulse-chase regime. Asterisks indicate non-specifically precipitated proteins.